Multidimensional Research Insight is an international scholarly journal of social sciences which provides wide, excellent cross-disciplinary research papers. It aims at increasing the generation of new self-integration knowledge with an emphasis on interdisciplinary research which harnesses interests cut across discipline to find solutions to local and global amplifications. The published journal is intended to enhance the probability of domain spanning and allow researchers to focus on the discovery of the connections on one field and others, and provide the overviews which are not partial.
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Background: The Siberian Tiger (Panthera tigris altaica) is a critically endangered keystone species. Accurate molecular identification of biological traces is essential for effective conservation, anti-poaching enforcement, and ecological monitoring. Objective: The main objective of the current study was to validate the performance of a cytb-targeted quantitative PCR assay for the molecular identification of Siberian Tiger DNA and to assess the utility of droplet digital PCR as a complementary confirmatory tool. Methods: The current study validated a standardized quantitative PCR (qPCR) assay targeting the mitochondrial cytochrome *b* (cytb) gene for specific detection of Siberian Tiger DNA. Assay performance was evaluated using the YouSeq Siberian Tiger qPCR Test Kit on known reference samples, non-target species (Arabian sand cat, cheetah), and serial dilutions. Confirmatory analysis was performed using droplet digital PCR (ddPCR) to ensure specificity and absolute quantification. Results: The qPCR assay demonstrated high sensitivity, with a limit of detection below 100 target copies per reaction, and high specificity, showing no cross-reactivity with non-target species. Siberian Tiger samples produced robust, early amplification (Cq 15.74–18.03). ddPCR provided absolute quantification, confirming target presence and resolving a case of ambiguous qPCR amplification from a White Tiger sample, highlighting ddPCR’s utility in eliminating false positives. Serial dilution analysis confirmed excellent dynamic range and reproducibility. Conclusion: This study validated a sensitive, specific, and reliable cytb-based qPCR/ddPCR assay for the molecular identification of Siberian Tiger DNA. The integrated approach provides a robust tool for wildlife forensics and conservation genetics, enabling accurate detection from trace and degraded samples to support legal enforcement and biodiversity monitoring.
Siberian Tiger; ; cytb gene; qPCR; ddPCR; wildlife forensics; conservation genetics; molecular identification.